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Literature summary for 1.1.1.85 extracted from

  • Suzuki, T.; Yasugi, M.; Arisaka, F.; Yamagishi, A.; Oshima, T.
    Adaptation of a thermophilic enzyme, 3-isopropylmalate dehydrogenase, to low temperatures (2001), Protein Eng., 14, 85-91.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli Thermus thermophilus

Protein Variants

Protein Variants Comment Organism
A31G/G43A/A709G optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°Cm increase in Km-values, slight loss of thermal stability Thermus thermophilus
C275T similar pH-dependence to that of the wild-type enzyme, improved binding affinities for both D-3-isopropylmalate and NAD+ result in an improved catalytic efficiency, mutant enzyme retains thermal stability Thermus thermophilus
G376A optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°C, increase in Km-values, mutant enzyme retains thermal stability Thermus thermophilus
G43A optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°C, increase ion Km-values, slight loss of thermal stability Thermus thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.001
-
isopropylmalate pH 8.0, 40°C, mutant enzyme C275T Thermus thermophilus
0.0012
-
isopropylmalate pH 8.0, 40°C, mutant enzyme G376A Thermus thermophilus
0.00125
-
isopropylmalate pH 8.0, 40°C, wild-type enzyme Thermus thermophilus
0.0013
-
isopropylmalate pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
0.0024
-
isopropylmalate pH 8.0, 40°C, mutant enzyme G43A Thermus thermophilus
0.0078
-
isopropylmalate pH 8.0, 70°C, mutant enzyme C275T Thermus thermophilus
0.0089
-
isopropylmalate pH 8.0, 70°C, wild-type enzyme Thermus thermophilus
0.0096
-
isopropylmalate pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
0.01
-
isopropylmalate pH 8.0, 70°C, mutant enzyme G376A Thermus thermophilus
0.011
-
isopropylmalate pH 8.0, 70°C, mutant enzyme G43A Thermus thermophilus
0.0149
-
NAD+ pH 8.0, 40°C, reaction with 3-D-isopropylmalate, mutant enzyme C275T Thermus thermophilus
0.0258
-
NAD+ pH 8.0, 40°C, isopropylmalate dehydrogenase wild-type Thermus thermophilus
0.157
-
NAD+ pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
0.16
-
NAD+ pH 8.0, 40°C, mutant enzyme G43A Thermus thermophilus
0.225
-
NAD+ pH 8.0, 40°C, isopropylmalate dehydrogenase cold-adapted mutant T1155 Thermus thermophilus
0.341
-
NAD+ pH 8.0, 70°C, isopropylmalate wild-type Thermus thermophilus
0.82
-
NAD+ at 70°C, mutant enzyme G43A Thermus thermophilus
0.917
-
NAD+ at 70°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.01
-
NAD+ at 40°C, isopropylmalate dehydrogenase cold-adapted mutant P215 Thermus thermophilus
3.56
-
NAD+ 70°C, isopropylmalate dehydrogenase cold-adapted mutant P215 Thermus thermophilus

Organism

Organism UniProt Comment Textmining
Thermus thermophilus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2R,3S)-3-isopropylmalate + NAD+
-
Thermus thermophilus 2-oxoisocaproate + NADH + H+ + CO2
-
?
(2R,3S)-3-isopropylmalate + NAD+
-
Thermus thermophilus (2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
r

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
82
-
10 min, 50% inactivation, mutant enzyme A31G/G43A/A709G Thermus thermophilus
85
-
10 min, 50% inactivation, mutant enzyme G43A Thermus thermophilus
87
-
10 min, 50% inactivation, wild-type enzyme, mutant enzyme C275T and mutant enzyme G376A Thermus thermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.36
-
NAD+ pH 8.0, 40°C, wild-type enzyme Thermus thermophilus
2.36
-
(2R,3S)-3-isopropylmalate pH 8.0, 40°C, wild-type enzyme Thermus thermophilus
2.41
-
NAD+ pH 8.0, 40°C, mutant enzyme C275T Thermus thermophilus
2.41
-
(2R,3S)-3-isopropylmalate pH 8.0, 40°C, mutant enzyme C275T Thermus thermophilus
7.2
-
NAD+ pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
7.2
-
(2R,3S)-3-isopropylmalate pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
10.7
-
NAD+ pH 8.0, 40°C, mutant enzyme G43A Thermus thermophilus
10.7
-
(2R,3S)-3-isopropylmalate pH 8.0, 40°C, mutant enzyme G43A Thermus thermophilus
17.7
-
NAD+ pH 8.0, 40°C, mutant enzyme G376A Thermus thermophilus
17.7
-
(2R,3S)-3-isopropylmalate pH 8.0, 40°C, mutant enzyme G376A Thermus thermophilus
44.7
-
NAD+ pH 8.0, 70°C, mutant enzyme C275T Thermus thermophilus
44.7
-
(2R,3S)-3-isopropylmalate pH 8.0, 70°C, mutant enzyme C275T Thermus thermophilus
45.8
-
NAD+ pH 8.0, 70°C, wild-type enzyme Thermus thermophilus
45.8
-
(2R,3S)-3-isopropylmalate pH 8.0, 70°C, wild-type enzyme Thermus thermophilus
50.1
-
NAD+ pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
50.1
-
(2R,3S)-3-isopropylmalate pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
63.6
-
NAD+ pH 8.0, 70°C, mutant enzyme G43A Thermus thermophilus
63.6
-
(2R,3S)-3-isopropylmalate pH 8.0, 70°C, mutant enzyme G43A Thermus thermophilus
100
-
NAD+ pH 8.0, 70°C, mutant enzyme G376A Thermus thermophilus
100
-
(2R,3S)-3-isopropylmalate pH 8.0, 70°C, mutant enzyme G376A Thermus thermophilus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Thermus thermophilus